KR20160068636A - Adjuvant composition comprising one or more influenza virus neutralizing binding molecules and vaccine composition comprising thereof - Google Patents

Abstract

The present invention relates to an adjuvant composition comprising one or more influenza virus neutralizing binding molecules and a vaccine composition comprising the same. Since the composition comprising one or more influenza virus neutralizing binding molecules of the present invention has been found to enhance the effect of the vaccine, it can be used as an adjuvant to increase the immune response upon administration of the vaccine, Very useful.

Description

[0001] The present invention relates to an adjuvant composition comprising one or more influenza virus neutralizing binding molecules and a vaccine composition comprising the influenza virus neutralizing binding molecules and vaccine composition,

The present invention relates to an adjuvant composition comprising one or more influenza virus neutralizing binding molecules and a vaccine composition comprising the same. More particularly, the present invention relates to a vaccine composition comprising an influenza virus that increases the vaccine-induced immune response, And to vaccine compositions comprising the same. BACKGROUND OF THE INVENTION < RTI ID = 0.0 > [0002] < / RTI >

Influenza virus (Influenza virus) is a disease that occurs in the respiratory tract, which is common in winter and is highly infectious and spreads easily to all ages, especially in the elderly (Treanor J, 2004, N Engl J Med ., 350 (3): 218-20). The enveloped virus belonging to Orthomyxoviridae has eight segments of negative-polar and single-strand RNA (ribonucleic acid) as the genome, and influenza virus The influenza A virus is divided into several subtypes according to HA (hemagglutinin) and NA (neuraminidase), which are major surface proteins, in the case of influenza A virus. To date, 17 types of HA and 10 types of NA have been known (Cheung TK and Poon LL 2007, Ann NY Acad Sci. 1102: 1-25; Tong S, et al. 2012, Proc. Natl. Acad. 109: 4269-4274). Influenza viruses continue to produce variant viruses due to the combination and mutation of various genes due to the nature of the virus being able to infect birds, swine and humans, and the genome of RNA segments (Treanor J, 2004. N Engl J Med 350 (3): 218-20). Because of this persistent mutation, it is difficult to obtain a permanent immunity. Therefore, the most effective preventive measures at present are vaccination against the influenza virus, which is expected to become popular every year, to form immunity to a specific type every year will be.

Currently, influenza virus vaccine, which is inoculated every year, is a trivalent or tetravalent vaccine, using HA of the influenza A H1 and H3 subtypes and one or two types of influenza B type HA.

Influenza virus vaccines and other infectious disease vaccines to increase the immunogenicity of substances to be added, which is called the adjuvant (adjuvant). Adjuvants approved for use in humans include Alum, which is composed of aluminum hydroxide and aluminum phosphate, and AS04, which consists of MF59, AS03 and TLR4 agonist MPL and aluminum hydroxide, with oil-in-water emulsions. (Rappuoli R, 2011. Nature Reviews Immunology 11 , 11 (12): 865-72)

In addition to the substances mentioned above, there are reports of increasing the immune response by administering the antibody together with the antigen, and attempts are being made to use the antibody as an adjuvant.

Antibodies against influenza A viruses patented by the present applicant have shown neutralizing efficacy against various influenza subtypes. In particular, the antibody disclosed in Korean Patent Application No. 10-2011-0020061 mainly comprises phylogenetic group 1 (H1, H2, H5, H9, etc.), the antibody disclosed in Korean Patent Application No. 10-2012-0107512 mainly exhibited neutralizing effect on phylogenetic group 2 (H3, H7, etc.). Therefore, a cocktail preparation which can prevent and treat all the viruses belonging to group 1 and group 2, which are likely to be prevalent by mixing two or more kinds of antibodies together, was developed, and Korean Patent Application No. 10-2014-0036601 Lt; / RTI >

The inventors of the present invention completed the present invention upon confirming that the effect of the vaccine can be enhanced by treating the influenza virus neutralizing antibody together when the vaccine is administered.

Accordingly, an object of the present invention is to provide an adjuvant composition comprising one or more influenza virus neutralizing binding molecules.

Another object of the present invention is to provide a vaccine composition comprising the adjuvant composition and the target antigen.

Another object of the present invention is to provide a method for preparing a vaccine by adding the adjuvant composition to a vaccine composition.

Another object of the present invention is to provide a method for enhancing the immunogenic activity of a vaccine composition by adding the adjuvant composition to the vaccine composition.

Another object of the present invention is to provide a method of immunizing a vaccine composition by administering the vaccine composition to a host.

Another object of the present invention is to provide a method for preparing an immunological product from an immunized host by administering the vaccine composition to a host.

In order to solve the above problems, the present invention provides an adjuvant composition comprising one or more influenza virus neutralizing binding molecules.

In one embodiment of the invention, the binding molecule can bind to an epitope in the stem region of the hemagglutinin (HA) protein of influenza A virus.

In one embodiment of the invention, the epitope of the first binding molecule may comprise amino acid residues at positions 18, 38, 40, 291, 292 and 318 of the HA1 polypeptide. In addition, the epitope of the first binding molecule may comprise amino acid residues at positions 18, 19, 20, 21, 41, 42, 45, 48, 49, 52 and 53 of the HA2 polypeptide.

In one embodiment of the invention, the epitope of the first binding molecule comprises the amino acid residues at positions 18, 38, 40, 291, 292 and 318 of the HA1 polypeptide and at positions 18, 19, 20, 21, 41, 42, 45, 48, 49, 52, and 53, respectively.

In one embodiment of the invention, the epitope of the second binding molecule may comprise amino acid residues at positions 278 and 318 of the HA1 polypeptide. In addition, the epitope of the second binding molecule may comprise amino acid residues at positions 38, 39, 41, 42, 45, 48, 49, 52, and 53 of the HA2 polypeptide. Also, the epitope of the second binding molecule comprises an amino acid residue at the position of the HA1 polypeptide and / or HA2 polypeptide of the HA first monomer and the position of the HA1 polypeptide of the second monomer adjacent to the first monomer at positions 25, 32, and 33 amino acid residues.

In one embodiment of the invention, the epitope of the second binding molecule comprises an amino acid residue at positions 278 and 318 of the HA1 polypeptide and at positions 38, 39, 41, 42, 45, 48, 49, 52, 53 amino acid residues. In another embodiment, the epitope of the second binding molecule comprises an HA1 polypeptide of the HA first monomer and an amino acid residue at the position of the HA2 polypeptide, wherein the epitope of the HA1 polypeptide of the second monomer adjacent to the first monomer Position 25, 32, and 33 amino acid residues.

In one embodiment of the invention, the epitope of the second binding molecule comprises amino acid residues at positions 278 and 318 of the HA1 polypeptide and at positions 38, 39, 41, 42, 45, 48, 49, 52, 53 , 58 and 99 amino acid residues. In another embodiment, the epitope of the second binding molecule comprises an HA1 polypeptide of the HA first monomer and an amino acid residue at the position of the HA2 polypeptide, wherein the epitope of the HA1 polypeptide of the second monomer adjacent to the first monomer Position 25, 27, 32 and 33 amino acid residues.

In one embodiment of the invention, the epitope of the second binding molecule comprises the amino acid residues at positions 54, 55, 278, 291 and 318 of the HA1 polypeptide and the positions 19, 20, 21, 38, 39, 41 , 42, 45, 46, 48, 49, 52, 53, 56, 57, and 60 amino acid residues. In another embodiment, the epitope of the second binding molecule comprises an HA1 polypeptide of the HA first monomer and an amino acid residue at the position of the HA2 polypeptide, wherein HA1 of the HA second monomer adjacent to the HA first monomer Amino acid residues at positions 25, 32, 33, 310, 311, and 312 of the polypeptide.

The amino acid position numbering of the epitope is based on H3 HA numbering.

The binding molecule of the present invention can inhibit the membrane fusion of virus and target cells. In addition, the binding molecule of the present invention can inhibit the virus by the Fc function of the antibody, that is, ADCC and CDC.

In one embodiment of the invention, the binding molecule comprises

According to the Kabat method

i) a light chain variable region comprising the CDR1 region of SEQ ID NO: 1, the CDR2 region of SEQ ID NO: 2, and the CDR3 region of SEQ ID NO: 3, and the CDR1 region of SEQ ID NO: 4, the CDR2 region of SEQ ID NO: 5, A binding molecule comprising a heavy chain variable region comprising a CDR3 region region; And

ii) a light chain variable region comprising the CDR1 region of SEQ ID NO: 7, the CDR2 region of SEQ ID NO: 8 and the CDR3 region of SEQ ID NO: 9, and the CDR1 region of SEQ ID NO: 10, the CDR2 region of SEQ ID NO: 11, A binding molecule comprising a heavy chain variable region comprising a CDR3 region

And a binding molecule selected from the group consisting of

In the present invention, the CDRs of the variable regions have been determined in a conventional manner according to a system designed by Kabat et al. (Kabat et al., Sequences of Proteins of Immunological Interest (5 ^th ), National Institutes of Health, Bethesda, MD (1991)). Although the CDR numbering used in the present invention uses the Kabat method, a binding molecule comprising a CDR determined according to other methods such as the IMGT method, Chothia method, AbM method and the like is also included in the present invention.

In another embodiment of the present invention, the binding molecule comprises

A binding molecule comprising a light chain comprising a polypeptide sequence of SEQ ID NO: 13 and a heavy chain comprising a polypeptide sequence of SEQ ID NO: 14; And

A light chain comprising a polypeptide sequence of SEQ ID NO: 15 and a binding molecule comprising a heavy chain comprising a polypeptide sequence of SEQ ID NO:

≪ RTI ID = 0.0 > and / or < / RTI >

The present invention also provides a vaccine composition comprising the adjuvant composition and the target antigen. Such target antigens may be, but are not limited to, viral antigens. Preferably, the viral antigen is an influenza virus antigen. The influenza virus antigen includes influenza A virus or influenza B virus antigen. The influenza virus antigen may be hemagglutinin (HA) or neuraminidase (NA), but is not limited thereto.

The present invention also provides a method for preparing a vaccine by adding the adjuvant composition to a vaccine composition. The vaccine composition may be, but is not limited to, an influenza virus vaccine composition.

The present invention also provides a method for enhancing the immunogenic activity of a vaccine composition by adding the adjuvant composition to the vaccine composition. The vaccine composition may be, but is not limited to, an influenza virus vaccine composition.

The present invention also provides a method for immunizing a vaccine composition by administering the vaccine composition to a host.

In addition,

a) administering the vaccine composition to a host to immunize; And

b) obtaining an immunological product from said immunized host

Lt; RTI ID = 0.0 > a < / RTI >

In addition, the present invention provides a method for preventing viral-derived diseases comprising administering to a subject an effective amount of a vaccine composition comprising the adjuvant composition. For example, the virus-derived disease may be an influenza virus-derived disease.

Hereinafter, terms used in the present invention are defined as follows.

"Influenza A virus" is an enveloped virus belonging to Orthomyxoviridae. It is an eight-segmented negative-sense, single-stranded RNA ribonucleic acid) as a genome. It is divided into A, B, and C groups and divided into several subtypes according to HA (hemaggutinin) and NA (neuraminidase). To date, 17 types of HA and 10 types of NA have been known.

The "H1 subtype" described in the present invention includes H1N1, H1N2, H1N3, H1N4, H1N5, H1N6, H1N7, H1N8, H1N9 and H1N10.

The "H2 subtype" described in the present invention includes H2N1, H2N2, H2N3, H2N4, H2N5, H2N6, H2N7, H2N8, H2N9 and H2N10.

The "H5 subtype" described in the present invention includes H5N1, H5N2, H5N3, H5N4, H5N5, H5N6, H5N7, H5N8, H5N9 and H5N10.

The "H9 subtype" described in the present invention includes H9N1, H9N2, H9N3, H9N4, H9N5, H9N6, H9N7, H9N8, H9N9 and H9N10.

The "H3 subtype" described in the present invention includes H3N1, H3N2, H3N3, H3N4, H3N5, H3N6, H3N7, H3N8, H3N9 and H3N10.

The "H7 subtype" described in the present invention includes H7N1, H7N2, H7N3, H7N4, H7N5, H7N6, H7N7, H7N8, H7N9 and H7N10.

"Hemagglutinin (hereinafter referred to as" HA ") refers to the envelope glycoprotein of influenza virus. HA mediates the influx of influenza viruses into the host cells. So far 17 types of subtypes have been reported.

"Neuraminidase (hereinafter referred to as" NA ") refers to an envelope protein of influenza virus. NA plays an important role when the influenza virus spreads and spreads. So far, 10 types of subtypes have been reported.

"Influenza vaccines" are the best way to prevent seasonal or pandemic influenza, largely divided into live vaccines and inactivated vaccines. Live attenuated vaccines have been developed and used. The inactivated vaccine is a split vaccine in which the virus envelope is pulverized with a whole virus vaccine or an ether that uses the whole virus in which the virus cultivated in the incubation column or cell culture is inactivated by formalin or the like, And a subunit vaccine in which the NA component is purified. A vaccine containing one type of influenza B group together with a H1 and H3 subtype of the influenza A group is called a trivalent vaccine and a vaccine containing two types of influenza B group is called a tetravalent vaccine.

The " influenza vaccine " described in the present invention includes both trivalent, tetravalent seasonal, live vaccine of pandemics and inactivated vaccine.

Binding molecule " as used herein refers to an intact immunoglobulin comprising a monoclonal antibody, such as a chimeric, humanized or human monoclonal antibody, or an immunoglobulin that binds to an antigen, for example, Receptor or protein capable of binding to a variable domain or substrate comprising an immunoglobulin fragment competing with intact immunoglobulin for binding to influenza A virus monomer HA or trimer HA. Regardless of structure, the antigen-binding fragments bind to the same antigen recognized by the intact immunoglobulin. The antigen-binding fragment may comprise two or more contiguous amino acid sequences of the binding molecule, at least 20 contiguous amino acid residues, at least 25 contiguous amino acid residues, at least 30 contiguous amino acid residues, at least 35 contiguous amino acid residues, , At least 50 contiguous amino acid residues, at least 60 contiguous amino acid residues, at least 70 contiguous amino acid residues, at least 80 contiguous amino acid residues, at least 90 contiguous amino acid residues, at least 100 contiguous amino acid residues, A peptide or polypeptide comprising an amino acid sequence of at least 150 consecutive amino acid residues, at least 175 consecutive amino acid residues, at least 200 consecutive amino acid residues, or at least 250 consecutive amino acid residues. "Antigen-binding fragments" specifically include Fab, F (ab '), F (ab') 2, Fv, dAb, Fd, complementarity determining region (CDR) fragments, single- Single chain antibodies, single-chain phage antibodies, diabodies, triabodies, tetrabodies, polypeptides containing one or more fragments of an immunoglobulin sufficient to bind to a particular antigen to a polypeptide, and the like. The fragment may be produced synthetically or by enzymatic or chemical degradation of the complete immunoglobulin, or may be genetically engineered by recombinant DNA technology. Methods of generation are well known in the art.

As used herein, the term " pharmaceutically acceptable excipient "means an inert material combined with an active molecule, such as a drug, agent, or binding molecule to produce an acceptable or convenient dosage form. Pharmaceutically acceptable excipients are non-toxic or at least those excipients which are acceptable for their intended use in the recipient at the dosages and concentrations for which they are used and include other components of the formulation including drugs, Can be compatible.

As used herein, the term " effective amount " refers to the amount of binding molecule of the invention that is effective in increasing the effect of a vaccine when administered with a vaccine of influenza A virus.

In the present invention, antibodies raised in the parent patent application (Korean Patent Application No. 10-2011-0020061, Korean Patent Application No. 10-2012-0107512, Korean Patent Application No. 10-2014-0036601) are used for influenza vaccination, Was confirmed in a mouse experiment. Korean Patent Application No. 10-2011-0020061, Korean Patent Application No. 10-2012-0107512, and Korean Patent Application No. 10-2014-0036601, which were filed by the present applicant, are incorporated herein by reference.

Since the composition comprising one or more influenza virus neutralizing binding molecules of the present invention has been found to enhance the effect of the vaccine, it can be used as an adjuvant to increase the immune response upon administration of the vaccine, Very useful.

Figure 1 shows the results of an ELISA to determine whether the antibody produced in the blood after 13, 17 and 27 days of administration of the vaccine and the CT120 antibody or the CT149 antibody or the representative Adjuvant (Alum) can detect the influenza virus.
FIG. 2 shows ELISA of the H1N1 HA protein or H1N1 influenza virus specific antibody in serum after the second injection of the H1N1 vaccine as shown in Table 3 and the serum was collected two weeks after the second injection.
Figure 3 is after a challenge with H1N1 influenza virus 10MLD _50, to confirm the survival of immune mouse with body weight variation.
FIG. 4 shows ELISA of H1N1 influenza virus specific antibody titers in serum after the injection of H1N1 vaccine and immunoadjuvant as shown in Table 5 and serum sampling after 2 weeks of intramuscular injection.
FIG. 5 shows ELISA of H1N1 HA protein or H1N1 influenza virus specific antibody in serum by taking H1N1 vaccine and immunoadjuvant as shown in Table 8 and taking serum at intervals of one week from the 2nd week after the first intramuscular injection.
Figure 6 after the challenge with H1N1 influenza virus 10MLD _50, to confirm the survival of immune mouse with body weight variation.
FIG. 7 shows ELISA of H1N1 HA protein or H1N1 influenza virus specific antibody in serum by taking H1N1 vaccine and immunoadjuvant as shown in Table 10 and taking serum at intervals of one week from the 2nd week after the first intramuscular injection.
Figure 8 after challenge the H1N1 influenza virus to 10MLD _50, to confirm the survival of immune mouse with body weight variation.

Hereinafter, the present invention will be described in detail with reference to examples. However, the following examples are intended to illustrate the contents of the present invention and are not intended to limit the scope of the invention. The documents cited in the present invention are incorporated herein by reference.

Example

Example  1: vaccine against influenza virus neutralizing antibodies Adjubant  Check the effect

1-1. ELISA   Experiment

Production of antibodies against influenza virus was confirmed in mice using 0.2 쨉 g of H1N1 split vaccine with CT120 antibody and CT149 antibody as an adjuvant of influenza virus vaccine.

At this time, alum (1 mg), which is a typical adjuvant, was used.

The specific design of the experiment is shown in the table below.

Influenza virus neutralizing antibody vaccine adjuvant effect confirmation experiment Group Antigen Adjuvant Route Mouse # Group 1 PBS - i.m. 4 Group 2 PBS CT 120 10ug i.m. 4 Group 3 PBS CT 149 10ug i.m. 4 Group 4 H1N1 split vaccine 0.2ug - i.m. 4 Group 5 H1N1 split vaccine 0.2ug CT 120 10ug i.m. 4 Group 6 H1N1 split vaccine 0.2ug CT 120 5ug i.m. 4 Group 7 H1N1 split vaccine 0.2ug CT 120 1ug i.m. 4 Group 8 H1N1 split vaccine 0.2ug CT 120 0.5ug i.m. 4 Group 9 H1N1 split vaccine 0.2ug CT 149 10ug i.m. 4 Group 10 H1N1 split vaccine 0.2ug CT 149 5ug i.m. 4 Group 11 H1N1 split vaccine 0.2ug CT 149 1ug i.m. 4 Group 12 H1N1 split vaccine 0.2ug CT 149 0.5ug i.m. 4 Group 13 H1N1 split vaccine 0.2ug Alum i.m. 4

After the experiment with the experimental design shown in Table 1, the mice were bled at 13 days, 17 days, and 27 days after administration of the vaccine and the adjuvant, and the vaccine administered and the antibody produced by the adjuvant detected influenza virus Were identified by ELISA. The results are shown in Fig.

After coating the plate with H1N1 virus, the amount of antibody capable of detecting the virus in the serum was checked. As a result, the CT120 or 149 or the group treated with Alum was equivalent to the vaccine as a whole It was confirmed that there was a large amount of antibodies capable of detecting viruses. Compared with the group treated with Alum, the group treated with 5 μg of CT120 and 10 μg of CT149 showed antibody production similar to that of the group treated with Alum, the group treated with 1 μg of CT149 and 0.5 μg of CT149 It was confirmed that more antibodies were produced compared to the group treated with Alum.

1-2. Hemagglutination Reaction Experiment HI assay , hemaglutinin inhibition assay )

The efficacy of the vaccine was evaluated by the HI assay and the hemaglutinin inhibition assay, which is one of the methods for evaluating the effectiveness of the vaccine. The HI assay uses the principle that an antibody capable of inhibiting hemagglutination hinders the binding of red blood cells to HA, the surface protein of influenza virus. Using the blood obtained on days 13, 17, and 27 after vaccination, we were able to determine how much antibody was produced to suppress the coagulation reaction of influenza virus and chicken erythrocytes (Table 2).

Hemagglutination assay (HI assay) Group
HI Titer Day 13 Day 17 Day 27 PBS N.D. N.D. N.D. 10 [mu] g CT120 N.D. N.D. N.D. 10 μg of CT149 N.D. N.D. N.D. Split 0.2 g N.D. N.D. 40 Split 0.2 g + CT120 10 g N.D. N.D. 40 Split 0.2 g + CT120 5 g N.D. N.D. 80 Split 0.2 g + CT120 1 g N.D. N.D. 40 Split 0.2 [mu] g + CT149 0.5 [ N.D. N.D. 40 Split 0.2 g + CT149 10 g N.D. N.D. 80 Split 0.2 g + CT149 5 g N.D. N.D. 40 Split 0.2 [mu] g + CT149 1 [ N.D. 40 160 Split 0.2 [mu] g + CT149 0.5 [ N.D. N.D. 160 Split 0.2 g + Alum N.D. 20 80

Similar to the results shown in Fig. 1, the group treated with 5 μg of CT120 and 10 μg of CT149 showed HI titer similar to that of the group treated with Alum 1 mg, compared with the group treated with Alum, The group treated with 1 ㎍ and 0.5 ㎍ showed more HI titers compared with the group treated with Alum.

When CT120 or CT149 were administered together with influenza virus vaccine, the immunogenicity was similar to or better than that of the adjuvant Alum, and CT120 and CT149 were found to be effective as adjuvants for influenza virus vaccine.

Example  2: H1N1  vaccine (cell-based) Antigen quantity  Animal experiments for crystals

Prior to testing the efficacy of adjuvants for therapeutic antibodies as in Table 3 below, animal experiments were performed to determine the appropriate dose of vaccine. As shown in the table below, the H1N1 vaccine was administered alone from 0.01 ug to 1 ug, or alum immunoadjuvant was administered, and the antibody titer was confirmed by ELISA and the neutralizing antibody titer was confirmed by HI. Standard was conducted to compare the experimental results with the trivalent vaccine being sold.

After intramuscular injection of the H1N1 vaccine into the mice, the immune response induced in each experimental group was confirmed. Two weeks after the second intramuscular injection, the serum was collected to confirm the antibody and neutralizing antibody response in the serum.

As shown in FIG. 2, the antibody titre was increased in proportion to the amount of the administered antigen, and it was confirmed that the antibody titer was generally higher when the alum immunoadjuvant was added. In addition, there was no significant difference between antibody titers to HA protein and antibody to H1N1 virus.

As shown in Table 4, it was confirmed that the HI titer was higher in the experimental group in which alum was added than in the experimental group containing only the antigen. In addition, when the antigen concentration was 0.1 ug and the antigen concentration was 0.05 ug, the HI titer was not confirmed when the antigen alone was added. However, when the immunosuppressant alum was added, the HI titer was increased 8 and 4 times, respectively.

Four weeks after the second immunization, 10MLD ₅₀ of CA / 04/09 H1N1 virus was infected through nasal inoculation and then the change in body weight and survival rate were confirmed for 15 days to confirm protective immunity against influenza virus.

As shown in FIG. 3, it was confirmed that the protective immunity effect against the influenza virus was high in the test group in which the immunoadjuvant was administered, compared with the test group administered with the antigen alone at all concentrations. In addition, in the experimental group of two antigen concentrations (0.1 ug and 0.05 ug) selected as preliminary candidates in the previous HI titer results, the survival rate of the 0.1 ug test group was 80% in the antigen-alone test group, In one experimental group, the survival rate was 100%. On the other hand, in the case of 0.05ug experimental group, the survival rate of the test group administered with the antigen alone was 60%, and the survival rate was 100% in the test group administered with the adjuvant.

The HI titer and the protective immunity against influenza virus were evaluated. The HI titer was different in the 0.1 ug group, but there was little difference according to the presence or absence of the immunosuppressant in the protective immunity effect. It was determined that 0.05 ug was the final antigen concentration and it was decided to carry out an experiment to confirm the effect of the adjuvant.

Example  3: H1N1  Animal experiments to determine the effect of immunoadjuvants of CT120 and CT149 using vaccine (cell-based) as an antigen

Based on the results of animal experiments to determine the vaccine dose, animal experiments for selected CT120 and CT149 therapeutic antibodies were planned as shown in Table 5.

CT120 (mIgG2a) and CT149 (mIgG2a), which are mouse forms of the selected therapeutic antibodies, are expected to be higher than CT120 or CT149 in animal studies. The amount of immunoadjuvant was adjusted to 4 doses by adding low concentration and high concentration based on 1: 1 (0.05 ug) compared with the amount of antigen.

The H1N1 vaccine was mixed with various concentrations of therapeutic antibodies and reacted at 37 ° C for 1 hour. After intramuscular injection into the mice, the immune response induced in each experimental group was confirmed. Two weeks after two intramuscular injections, the serum was collected and the antibody and neutralizing antibody response were confirmed.

As shown in Fig. 4, the antibody titer showed the highest antibody titer at different concentrations depending on the kinds of therapeutic antibodies added together. And the antibody titer was markedly increased when it was injected 2 times. However, the difference in antibody titers did not show any significant difference in all experimental groups, and the experimental results were derived by synthesizing HI titer and influenza defense immunity.

Similar to the results of previous vaccine dose tests, as shown in Table 4, 0.05 μg, 0.1 μg, and 0.5 μg of CT149 (mIgG2a) were administered simultaneously with 0.05 μg of CT149 and no HI titer was observed in the antigen- In the experimental group, the HI titer was increased.

The HI titer of CTU20 (mIgG2a) was also increased by 0.05 ug and 0.1 ug, respectively. The HI titer was also increased in the experimental group with CT120 (mIgG2a).

The HI titer was not significantly different, so we could not elucidate the effect of HI titer. In addition, we performed an experiment to confirm the protective immunity effect against influenza virus.

In order to confirm protective immunity against influenza virus, 10 weeks after the second immunization, 10MLD ₅₀ of CA / 04/09 H1N1 virus was infected through nasal inoculation and survival was observed for 15 days thereafter.

As shown in Table 7, it was confirmed that the survival rate of mouse was generally increased in the experimental group in which the therapeutic antibody was added at an appropriate concentration as compared with the test group administered with the antigen alone. However, in the case of 0.1ug of CT149 (mIgG2a), the HI titer was higher than that of the antigen alone, but the protective immunity effect was not different.

CT149 showed the lowest defense immunity among the four therapeutic antibodies compared with the experimental results. The survival rate of CT149 was 50% at 0.5ug of CT149, the highest survival rate. CT149 (mIgG2a) showed a higher survival rate than CT149 as an immunosuppressive agent. In particular, the survival rate of mice treated with 0.01 ug and 0.05 ug was increased to 75%. CT120 showed the highest survival rate among the four therapeutic antibodies. In particular, 0.05 ug of the antibody showed the same HI titer and 100% survival rate as alum. It was also confirmed that the survival rate of CTU20 (mIgG2a) was also 75% in the experimental group containing 0.1 ug and 0.5 ug.

Example  4: H1N1  A large-scale animal experiment to confirm the effect of CT120 immunoadjuvant using vaccine (cell-based) as an antigen

Based on the above experimental results, we selected three concentrations of each of the four therapeutic antibodies as immunosuppressive agents, and increased the number of mice per experimental group to conduct a large-scale animal experiment.

As shown in Table 8, for each of CT120 and CT120 (mIgG2a), three concentrations were selected from the preliminary experiments, and 10 mice were set per experiment group. Unlike the previous experiment, serum was collected every week after immunization to confirm the change of antibody titer in serum and change of HI titer.

As shown in Fig. 5, the antibody titer was higher in the test group using mouse form CT120 (mIgG2a) as an immunizing adjuvant than in the test group using CT120 as an immunizing adjuvant. In the test group using CT120 (mIgG2a), the antibody titer was generally higher than the test group using alum as an immunosuppressant, and the antibody titer rapidly increased in the serum collected one week after the second immunization.

As shown in Table 9, the HI titer was increased up to 4 times according to the concentration in the test group in which CT120 (mIgG2a) was used as an immunosuppressant compared to the test group in which CT120 was used as an immunosuppressant. In the HI titer group, the highest HI titer was obtained in the test group using 0.5 ug of CT120 (mIgG2a), which was highest in the antibody titers.

Four weeks after the second immunization, 10MLD ₅₀ of CA / 04/09 H1N1 virus was infected through nasal inoculation and then survival was checked for 15 days to confirm protective immunity against influenza virus.

As shown in FIG. 6, when CT120 (mIgG2a) was used as an immunosuppressant, the survival rate was higher than that of CT120 as an immunizing adjuvant or antigen alone, and weight change was the least. In particular, the survival rate was 100% in the test group using 0.5ug of CT120 (mIgG2a), which is the highest level of antibody titer and HI titer, and the survival rate was 30% higher than that of the test group using alum as the immunosuppressant.

Based on these experimental results, it was confirmed that CT120 (mIgG2a) was more effective as an immunosuppressive agent than CT120, and it was confirmed that the effect was the best when treated at a concentration of 0.5 ug.

Example  5: H1N1  Large-scale animal studies to confirm the effect of CT149 immunoadjuvant using vaccine (cell-based) as an antigen

Similarly, large-scale animal experiments were conducted on CT149. As shown in Table 10, when CT149 was also confirmed through preliminary experiments, the experiment was conducted by selecting three concentrations that were the most effective.

As shown in Fig. 7, unlike CT120, the immunoadjuvant effect of CT149 was generally weak, and in the experimental group using CT149 (mIgG2a) 0.5g, antibody titers The synergistic effect could not be confirmed.

As shown in Table 11, in the test group in which CT149 or CT149 (mIgG2a) was used as an immunosuppressant, the HI titer did not increase significantly, and in the test group using CTU99 (mIgG2a), which was slightly higher in the antibody titer, The same antibody titers were observed.

In order to confirm protective immunity against influenza virus, 10 weeks after the second immunization, 10MLD ₅₀ of CA / 04/09 H1N1 virus was infected through nasal inoculation and survival was observed for 15 days thereafter.

As shown in Fig. 8, in CT149 and CT149 (mIgG2a), the survival rate of CT149 (mIgG2a) as an immunosuppressive agent was 80%, which was 10% higher than that of alum No immunosuppressant effect was observed. Weight change did not improve significantly compared to alum.

Taken together, these results indicate that CT120 (mIgG2a) was the most effective adjuvant for the H1N1 vaccine, and 0.5ug was the most effective. In the case of CT149 (mIgG2a), the effect of 0.5ug was shown to be lower than that of CT120 (mIgG2a).

Example  6: H3N2  vaccine (cell-based) Antigen quantity  Animal experiments for crystals

The effect of CT120 and CT149 on the H1N1 vaccine as an immunosuppressive agent was confirmed by various experiments and the H3N2 vaccine was produced using another influenza virus strain, Philippines / 2/82 (H3N2) Animal experiments were conducted to confirm the effect of CT120 and CT149 immunoadjuvants.

As shown in Table 12 below, prior to testing the efficacy of the therapeutic adjuvant, animal experiments were conducted to determine the appropriate vaccine dose. As in the table below, the H3N2 vaccine was administered alone or in combination with alum immunoadjuvant from 0.01 ug to 1 ug, and the antibody titer was confirmed by ELISA and the neutralizing antibody titer was confirmed by HI. In order to confirm the defense immunity effect, survival rate and body weight change were also confirmed by infection with mouse adapted Philippines / 2/82 (H3N2) virus.

<110> CELLTRION INC. <120> Adjuvant composition comprising one or more influenza virus          neutralizing binding molecules and vaccine composition comprising          the <130> CPD2014032KR <160> 16 <170> Kopatentin 2.0 <210> 1 <211> 11 <212> PRT <213> Artificial Sequence <220> <223> CT120 light chain CDR1 <400> 1 Arg Ala Ser Glu Asn Ile Trp Asn Asn Leu Ala   1 5 10 <210> 2 <211> 7 <212> PRT <213> Artificial Sequence <220> <223> CT120 light chain CDR2 <400> 2 Gly Ala Ser Thr Gly Ala Thr   1 5 <210> 3 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> CT120 light chain CDR3 <400> 3 Gln Gln Tyr Asn Ser Trp Pro Arg Thr   1 5 <210> 4 <211> 5 <212> PRT <213> Artificial Sequence <220> <223> CT120 heavy chain CDR1 <400> 4 Ser His Ala Ile Ser   1 5 <210> 5 <211> 17 <212> PRT <213> Artificial Sequence <220> <223> CT120 heavy chain CDR2 <400> 5 Gly Ile Ser Pro Met Phe Gly Thr Thr His Tyr Ala Gln Lys Phe Gln   1 5 10 15 Gly     <210> 6 <211> 14 <212> PRT <213> Artificial Sequence <220> <223> CT120 heavy chain CDR3 <400> 6 Asp Gly Ala Gly Ser Tyr Tyr Pro Leu Asn Trp Phe Asp Pro   1 5 10 <210> 7 <211> 12 <212> PRT <213> Artificial Sequence <220> <223> CT149 light chain CDR1 <400> 7 Arg Ala Ser His Arg Val Gly Ser Thr Tyr Ile Ala   1 5 10 <210> 8 <211> 7 <212> PRT <213> Artificial Sequence <220> <223> CT149 light chain CDR2 <400> 8 Gly Ala Ser Asn Arg Ala Thr   1 5 <210> 9 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> CT149 light chain CDR3 <400> 9 Gln Gln Phe Ser Val Ser Pro Trp Thr   1 5 <210> 10 <211> 5 <212> PRT <213> Artificial Sequence <220> <223> CT149 heavy chain CDR1 <400> 10 Thr Tyr Gly Val Ser   1 5 <210> 11 <211> 17 <212> PRT <213> Artificial Sequence <220> <223> CT149 heavy chain CDR2 <400> 11 Trp Ile Ser Ala Tyr Thr Gly Ile Thr Asp Tyr Ala Gln Lys Phe Gln   1 5 10 15 Gly     <210> 12 <211> 17 <212> PRT <213> Artificial Sequence <220> <223> CT149 heavy chain CDR3 <400> 12 Asp Lys Val Gln Gly Arg Val Glu Val Gly Ser Gly Gly Arg His Asp   1 5 10 15 Tyr     <210> 13 <211> 234 <212> PRT <213> Artificial Sequence <220> <223> CT120 light chain <400> 13 Met Glu Ala Pro Ala Gln Leu Leu Phe Leu Leu Leu Leu Trp Leu Pro   1 5 10 15 Asp Thr Thr Gly Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser              20 25 30 Leu Ser Pro Gly Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Glu Asn          35 40 45 Ile Trp Asn Asn Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro      50 55 60 Arg Leu Leu Ile Ser Gly Ala Ser Thr Gly Ala Thr Gly Val Ser Ser  65 70 75 80 Arg Phe Arg Gly Ser Gly Ser Arg Thr Glu Phe Thr Leu Thr Ile Ser                  85 90 95 Ser Leu Gln Ser Glu Asp Phe Ala Ile Tyr Phe Cys Gln Gln Tyr Asn             100 105 110 Ser Trp Pro Arg Thr Phe Gly Pro Gly Thr Lys Val Glu Ile Lys Arg         115 120 125 Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln     130 135 140 Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr 145 150 155 160 Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser                 165 170 175 Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr             180 185 190 Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys         195 200 205 His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro     210 215 220 Val Thr Lys Ser Phe Asn Arg Gly Glu Cys 225 230 <210> 14 <211> 472 <212> PRT <213> Artificial Sequence <220> <223> CT120 heavy chain <400> 14 Met Asp Trp Thr Trp Arg Phe Leu Phe Val Val Ala Ala Ala Thr Gly   1 5 10 15 Val Gln Cys Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Met              20 25 30 Pro Gly Ser Ser Val Lys Val Ser Cys Lys Thr Ser Gly Val Phe Phe          35 40 45 Ser Ser His Ala Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu      50 55 60 Glu Trp Met Gly Gly Ile Ser Pro Met Phe Gly Thr Thr His Tyr Ala  65 70 75 80 Gln Lys Phe Gln Gly Arg Val Thr Ile Thr Ala Asp Gln Ser Thr Thr                  85 90 95 Thr Ala Tyr Met Glu Leu Thr Ser Leu Thr Ser Glu Asp Thr Ala Val             100 105 110 Tyr Tyr Cys Ala Arg Asp Gly Ala Gly Ser Tyr Tyr Pro Leu Asn Trp         115 120 125 Phe Asp Pro Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ser Ser     130 135 140 Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr 145 150 155 160 Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro                 165 170 175 Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val             180 185 190 His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser         195 200 205 Ser Val Val Thr Val Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile     210 215 220 Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Arg Val 225 230 235 240 Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala                 245 250 255 Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro             260 265 270 Lys Asp Thr Leu Met Ile Ser Arg Thr Arg Glu Val Thr Cys Val Val         275 280 285 Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val     290 295 300 Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln 305 310 315 320 Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln                 325 330 335 Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala             340 345 350 Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro         355 360 365 Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr     370 375 380 Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser 385 390 395 400 Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr                 405 410 415 Lys Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr             420 425 430 Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe         435 440 445 Ser Cys Ser Val Met His Glu Gly Leu His Asn His Tyr Thr Gln Lys     450 455 460 Ser Leu Ser Leu Phe Pro Gly Lys 465 470 <210> 15 <211> 215 <212> PRT <213> Artificial Sequence <220> <223> CT149 light chain <400> 15 Glu Val Val Leu Thr Gln Ser Pro Gly Thr Leu Ala Leu Pro Pro Gly   1 5 10 15 Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser His Arg Val Gly Ser Thr              20 25 30 Tyr Ile Ala Trp Tyr Gln Gln Lys Ser Gly Gln Ala Pro Arg Arg Leu          35 40 45 Ile Tyr Gly Ala Ser Asn Arg Ala Thr Asp Ile Pro Asp Arg Phe Ser      50 55 60 Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Arg Arg Leu Glu  65 70 75 80 Pro Glu Asp Ser Ala Val Tyr Tyr Cys Gln Gln Phe Ser Val Ser Pro                  85 90 95 Trp Thr Phe Gly Gln Gly Thr Arg Val Glu Ile Lys Arg Thr Val Ala             100 105 110 Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser         115 120 125 Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu     130 135 140 Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser 145 150 155 160 Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu                 165 170 175 Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val             180 185 190 Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys         195 200 205 Ser Phe Asn Arg Gly Glu Cys     210 215 <210> 16 <211> 456 <212> PRT <213> Artificial Sequence <220> <223> CT149 heavy chain <400> 16 Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala   1 5 10 15 Ser Val Lys Val Ser Cys Lys Thr Ser Gly Tyr Ser Phe Ser Thr Tyr              20 25 30 Gly Val Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Pro Glu Trp Val          35 40 45 Gly Trp Ile Ser Ala Tyr Thr Gly Ile Thr Asp Tyr Ala Gln Lys Phe      50 55 60 Gln Gly Arg Val Thr Leu Thr Thr Asp Ala Thr Thr Ala Thr Ala Phe  65 70 75 80 Leu Asp Leu Arg Ser Leu Arg Pro Asp Asp Thr Ala Thr Tyr Phe Cys                  85 90 95 Ala Arg Asp Lys Val Gln Gly Arg Val Glu Val Gly Ser Gly Gly Arg             100 105 110 His Asp Tyr Trp Gly Gln Gly Thr Leu Val Ile Val Ser Ser Ala Ser         115 120 125 Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr     130 135 140 Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro 145 150 155 160 Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val                 165 170 175 His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser             180 185 190 Ser Val Val Thr Val Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile         195 200 205 Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val     210 215 220 Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala 225 230 235 240 Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro                 245 250 255 Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val             260 265 270 Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val         275 280 285 Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln     290 295 300 Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln 305 310 315 320 Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala                 325 330 335 Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro             340 345 350 Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr         355 360 365 Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser     370 375 380 Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr 385 390 395 400 Lys Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr                 405 410 415 Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe             420 425 430 Ser Cys Ser Val Met His Glu Gly Leu His Asn His Tyr Thr Gln Lys         435 440 445 Ser Leu Ser Leu Ser Pro Gly Lys     450 455

Claims (21)

An adjuvant composition comprising one or more influenza virus neutralizing binding molecules.
The method according to claim 1,
Wherein said binding molecule binds to an epitope in the stem region of the hemagglutinin (HA) protein of influenza A virus.
3. The method of claim 2,
Wherein said binding molecule is capable of neutralizing one or more influenza A virus subtypes selected from the group consisting of H1, H2, H5 and H9.
3. The method of claim 2,
Wherein the binding molecule is capable of neutralizing one or more influenza A virus subtypes selected from the group consisting of H1, H3, H5, H7 and H9.
The method of claim 3,
The epitope of the binding molecule is
The amino acid residues at positions 18, 38, 40, 291, 292 and 318 of the HA1 polypeptide,
An amino acid residue at position 18, 19, 20, 21, 41, 42, 45, 48, 49, 52 and 53 of the HA2 polypeptide.
5. The method of claim 4,
The epitope of the binding molecule is
The amino acid residues at positions 278 and 318 of the HA1 polypeptide,
39, 41, 42, 45, 48, 49, 52, and 53 of the HA2 polypeptide.
The method according to claim 6,
Wherein the epitope of the binding molecule comprises the HA1 polypeptide of the HA first monomer and the amino acid residue at the position of the HA2 polypeptide,
And an amino acid residue at positions 25, 32 and 33 of the HA1 polypeptide of the HA second monomer adjacent to the HA first monomer.
The method according to claim 6,
The epitope of the binding molecule is
Lt; RTI ID = 0.0 &gt; 58 &lt; / RTI &gt; and 99 amino acid residues of HA2 polypeptide.
8. The method of claim 7,
Wherein the epitope of the binding molecule further comprises a 27 amino acid residue at the position of the HA1 polypeptide of the second monomer adjacent to the first monomer.
The method according to claim 6,
The epitope of the binding molecule is
An amino acid residue at positions 54, 55 and 291 of the HA1 polypeptide and an amino acid residue at positions 19, 20, 21, 46, 56, 57 and 60 of the HA2 polypeptide.
8. The method of claim 7,
Wherein the epitope of the binding molecule further comprises amino acid residues at positions 310, 311, and 312 of the HA1 polypeptide of the HA second monomer adjacent to the HA first monomer.
The method according to claim 1,
The at least one binding molecule
i) a light chain variable region comprising the CDR1 region of SEQ ID NO: 1, the CDR2 region of SEQ ID NO: 2, and the CDR3 region of SEQ ID NO: 3 according to the Kabat method and the CDR1 region of SEQ ID NO: 4, A CDR2 region of SEQ ID NO: 6, and a CDR3 region region of SEQ ID NO: 6; And
ii) a light chain variable region comprising the CDR1 region of SEQ ID NO: 7, the CDR2 region of SEQ ID NO: 8, and the CDR3 region of SEQ ID NO: 9 according to the Kabat method and the CDR1 region of SEQ ID NO: 10, And a heavy chain variable region comprising the CDR3 region of SEQ ID NO: 12,
&Lt; / RTI &gt; wherein the binding molecule is one or more binding molecules selected from the group consisting of: &lt; RTI ID = 0.0 &gt;
The method according to claim 1,
The at least one binding molecule
A binding molecule comprising a light chain comprising a polypeptide sequence of SEQ ID NO: 13 and a heavy chain comprising a polypeptide sequence of SEQ ID NO: 14; And
A light chain comprising a polypeptide sequence of SEQ ID NO: 15 and a binding molecule comprising a heavy chain comprising a polypeptide sequence of SEQ ID NO:
&Lt; / RTI &gt; wherein the binding molecule is one or more binding molecules selected from the group consisting of: &lt; RTI ID = 0.0 &gt;
14. A vaccine composition comprising an adjuvant composition according to any one of claims 1 to 13 and a target antigen.
15. The method of claim 14,
Wherein the target antigen is an influenza virus antigen.
16. The method of claim 15,
Wherein the influenza virus antigen is hemagglutinin (HA) or neuraminidase (NA).
14. A method of preparing a vaccine by adding the adjuvant composition of any one of claims 1 to 13 to the vaccine composition.
14. A method of increasing the immunogenic activity of a vaccine composition by adding the adjuvant composition of any one of claims 1 to 13 to the vaccine composition.
14. A method of immunizing a vaccine composition of claim 14 by administering to the host.
a) administering to the host a vaccine composition of claim 14 to immunize; And
b) obtaining an immunological product from said immunized host
RTI ID = 0.0 &gt; 1, &lt; / RTI &gt;
21. The method of claim 20,
Wherein said immunological product is a T cell, a B cell, or an antibody.

Images